Characterization of antimicrobial resistance genes and their genetic context in Klebsiella spp. producers of KPC isolated from hospitals in the State of São Paulo ; Caracterização de genes de resistência antimicrobiana e seu contexto genético em Klebsiella spp. produtoras de KPC isoladas de hospitais do Estado de São Paulo

Klebsiella pneumoniae producing ESBLs are frequently involved in hospital infections in intensivecare unit (ICU). ESBLs are capable of hydrolyzing all penicillins, cephalosporins and aztreonam.Carbapenems are considered the agents of choice for treatment of infections caused by ESBLproducing microorganisms. However, Klebsiella spp. producing carbapenemases has become morefrequent. Moreover, K. pneumoniae producing 16S RMTase, RmtD, which confers high levelresistance to aminoglycosides, have been described in Brazil, limiting the therapeutical options.The aim of this study was to characterize resistance genes responsible to the production of ESBL,carbapenemases and 16S RMTases and their genetic context in Klebsiella spp. producing KPCfrom hospitals at Sao Paulo State. A hundred K. pneumoniae strains isolated in the period of 2009and 2011 and previously confirmed as KPC producers were subjected to PCR for the detectionof blaSHV, blaTEM, blaCTX-M, blaCTX-M-15, blaCTX-M-2, blaCTX-M-8, blaGES-1, blaVIM,blaIMP, blaSPM, blaNDM, blaOXA-48, armA, rmtA, rmtB rmtC e rmtD and DNA sequencing.Identification of Klebsiella spp. by biochemical tests, detection of blaSHV, blaLEN e blaOKPgenes by Multiplex PCR and 16S rRNA gene and rpoB sequencing was performed on blaSHVnegative isolates. Transformation and genomic cloning were performed for the identification ofa new aminoglycosides resistance gene. Sequencing of plasmids carrying 16S RMTases geneswas performed. All strains showed high resistance levels for most antibiotics tested. Our resultsshowed a positivity of 80% for blaCTX-M (61 CTX-M-15, 7 CTX-M-2, 2, 7%7 CTX- M-8 and 1CTX-M-35). Co-production of CTX-M-2 e CTX-M-15 was observed in two strains and CTX-M-8and CTX-M-59 co-production were also found in two strains. The presence of blaTEM-1 wasobserved in 26% of the strains. blaSHV were observed in 97% of the strains (89 SHV-11, 1 SHV1, 3 SHV-110 and 4 SHV-12). All Klebsiella spp. were characterized as KPC-2 producers. Of thethree blaSHV negative strains, 2 were confirmed as ...